USP19 Stabilizes TAK1 to Regulate High Glucose/Free Fatty Acid-induced Dysfunction in HK-2 Cells

• Published in: Current medical science Vol. 44; no. 4; pp. 707 - 717
• Main Authors: Yan, Xiao-hui, Zhu, Yin-na, Zhu, Yan-ting
• Format: Journal Article
• Language: English
• Published: Wuhan Huazhong University of Science and Technology 01.08.2024
• Subjects: Apoptosis - drug effects; Cell Line; Cell Proliferation - drug effects; Diabetic Nephropathies - genetics; Diabetic Nephropathies - metabolism; Endopeptidases - genetics; Endopeptidases - metabolism; Fatty Acids, Nonesterified - adverse effects; Fatty Acids, Nonesterified - metabolism; Fatty Acids, Nonesterified - pharmacology; Glucose - pharmacology; Humans; MAP Kinase Kinase Kinases - genetics; MAP Kinase Kinase Kinases - metabolism; Medicine; Medicine & Public Health; Original Article; Protein Kinases; Ubiquitination - drug effects; deubiquitination; high glucose; free fatty acid; USP19; HK-2 cells; dysfunction
• ISSN: 2096-5230; 2523-899X; 2523-899X
• DOI: 10.1007/s11596-024-2906-y

Objective Obesity-induced kidney injury contributes to the development of diabetic nephropathy (DN). Here, we identified the functions of ubiquitin-specific peptidase 19 (USP19) in HK-2 cells exposed to a combination of high glucose (HG) and free fatty acid (FFA) and determined its association with TGF-beta-activated kinase 1 (TAK1). Methods HK-2 cells were exposed to a combination of HG and FFA. USP19 mRNA expression was detected by quantitative RT-PCR (qRT-PCR), and protein analysis was performed by immunoblotting (IB). Cell growth was assessed by Cell Counting Kit-8 (CCK-8) viability and 5-ethynyl-2′-deoxyuridine (EdU) proliferation assays. Cell cycle distribution and apoptosis were detected by flow cytometry. The USP19/TAK1 interaction and ubiquitinated TAK1 levels were assayed by coimmunoprecipitation (Co-IP) assays and IB. Results In HG+FFA-challenged HK-2 cells, USP19 was highly expressed. USP19 knockdown attenuated HG+FFA-triggered growth inhibition and apoptosis promotion in HK-2 cells. Moreover, USP19 knockdown alleviated HG+FFA-mediated PTEN-induced putative kinase 1 (PINK1)/Parkin pathway inactivation and increased mitochondrial reactive oxygen species (ROS) generation in HK-2 cells. Mechanistically, USP19 stabilized the TAK1 protein through deubiquitination. Importantly, increased TAK1 expression reversed the USP19 knockdown-mediated phenotypic changes and PINK1/Parkin pathway activation in HG+FFA-challenged HK-2 cells. Conclusion The findings revealed that USP19 plays a crucial role in promoting HK-2 cell dysfunction induced by combined stimulation with HG and FFAs by stabilizing TAK1, providing a potential therapeutic strategy for combating DN.